CN106613934A - Method for rapidly propagating apple rootstock SH6 - Google Patents
Method for rapidly propagating apple rootstock SH6 Download PDFInfo
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- CN106613934A CN106613934A CN201510713945.2A CN201510713945A CN106613934A CN 106613934 A CN106613934 A CN 106613934A CN 201510713945 A CN201510713945 A CN 201510713945A CN 106613934 A CN106613934 A CN 106613934A
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Abstract
The invention provides a method for rapidly propagating apple rootstock SH6. The method comprises the steps of primary culture, propagation expanding sub-culture, rooting culture, seedling hardening and field transplanting to rapid propagate the apple rootstock SH6. The method for rapidly propagating the apple rootstock SH6 has the advantages of high value-added index, short rooting time and high transplanting survival rate, and has contribution to realization of tissue culture factory propagation of a dwarf apple rootstock variety SH6.
Description
Technical field
The invention belongs to the field of tissue culture of plant, and in particular to a kind of quick breeding apple anvil
The method of wooden SH6.
Background technology
Apple Dwarf Stocks planting type is paid much attention in the world for a long time, the great majority such as America and Europe
National more than 90% Apple Culture adopts short anvil planting type, and short anvil cultivation is also China's apple from now on
The developing direction of fruit production.SH6 is the Dwarf Stocks For Apple Trees that China cultivates, with resistance
By force, result is early, yield is high and superior quality feature, due to it from take root difficulty the problems such as, in bag
Include Beijing still to plant in dwarfing interstock mode in interior China's Main Apple Varieties producing region.Using plant
Tissue culture technique can at short notice obtain substantial amounts of plantlet in vitro, at present in species such as flowers
More practical application successful case has been obtained, because it has not by time and external environment bar
Part control, Economization on land, the advantage of quickness and high efficiency, it has also become a kind of wide variety of nursery hand
Section, contributes to accomplishing scale production.But in prior art, Dwarf Stocks For Apple Trees kind SH6
Plantlet in vitro squamous subculture growth coefficient is low, and adventitious root occurs difficulty, and field-transplanting survival rate is relatively low,
Limit its application in tissue culture industrial seedling rearing.
The content of the invention
For growth coefficient present in Dwarf Stocks For Apple Trees kind SH6 tissue culture is low, adventitious root
There is the low present situation of difficult, field-transplanting survival rate, the present invention provides a kind of quick breeding apple
The method of stock SH6, comprises the steps：
1) just generation and squamous subculture：By the sterilized sterilization of apple rootstock SH6 containing single leaf
Leaf stem section successively carries out Initial culture and expands numerous subculture training with Initial culture base and subculture medium
Support；
2) culture of rootage：Cut step 1) culture after the stem section containing simple bud, be inoculated into life
Culture of rootage is carried out on root culture medium；
3) hardening and field-transplanting：When the plantlet in vitro root length of culture of rootage reaches 2～3 centimetres,
Carry out hardening and field-transplanting；
The Initial culture base is with MS solid mediums as minimal medium, containing 6- benzyl ammonia
It is 0.1～0.6mg/L, poly- that the concentration of base purine is 0.1～0.8mg/L, the concentration of indolebutyric acid
Vinylpyrrolidone concentration be 200～400mg/L, sucrose concentration be 25～30g/L, agarose
Concentration be 5～7g/L culture medium；
The subculture medium is with MS solid mediums as minimal medium, containing 6- benzyl ammonia
The concentration of base purine be 0.1～0.8mg/L, indolebutyric acid concentration be 0.1～0.6mg/L, sugarcane
The concentration of sugar is 25～30g/L, the culture medium that the concentration of agarose is 5～7g/L；
The root media is with 1/2MS solid mediums as minimal medium, containing Yin
The concentration of diindyl butyric acid be 0.1～1.2mg/L, the concentration of sucrose be 15～25g/L, agarose it is dense
Spend the culture medium for 5～7g/L.
Wherein, the Initial culture base, polyvinylpyrrolidoneconcentration concentration is preferably 400mg/L.
Wherein, the root media, the concentration of indolebutyric acid is preferably 0.5～1.0mg/L,
More preferably 0.7～0.9mg/L, more preferably 0.7～0.8mg/L, most preferably
0.8mg/L。
Wherein, step 1) sterilization, concrete grammar is：Rinsed 30 minutes with water,
The alcohol disinfecting of concentration expressed in percentage by volume 70% 10～60 seconds, aseptic water washing 2～4 times, quality
The mercuric chloride of percentage concentration 0.1% is sterilized 3～10 minutes, aseptic water washing 2～4 times.
Wherein, the Initial culture and numerous squamous subculture is expanded, condition of culture is, temperature 20-28 DEG C,
Humidity 55-75%, hour/day of illumination 14～18, intensity of illumination 1800-2200Lx.
Wherein, the culture of rootage, is divided into two stages：
The light culture stage：6～12 days, temperature 18-26 DEG C, humidity 55-75%；
The optical culture stage：6～10 days, temperature 18-26 DEG C, humidity 55-75%, illumination 16～20
Hour/day, intensity of illumination 1800-2200Lx.
Wherein, the hardening, is bottle hardening to be closed in warmhouse booth 3 days, corkage hardening 3 days.
Wherein, the field-transplanting, to wash away the culture medium of plantlet in vitro of taking root, using quality hundred
The carbendazim of point concentration 0.2% carries out root sterilization, is transplanted into peat：Perlite：Vermiculite body
Product is than being 1：1：In 1 matrix, after root system development is complete land for growing field crops is transplanted to.
The method that the present invention also provides described quick breeding apple rootstock SH6, it is numerous in apple
Application in educating.
The beneficial effects of the present invention is：
The present invention can realize the quick breeding by group culture of Dwarf Stocks For Apple Trees SH6, squamous subculture
During 25 days cycles, value-added coefficient reaches 8～10 times, and rootage duration is no longer than 15 days, takes root
Rate reaches more than 90%, and field-transplanting survival rate is more than 85% after hardening domestication, is Apple Dwarf
Changing stock variety SH6 tissue cultures plant modification provides simple and fast efficient technical method.
Description of the drawings
Fig. 1 is Dwarf Stocks For Apple Trees SH6 squamous subculture photos in embodiment 1；
Fig. 2 is Dwarf Stocks For Apple Trees SH6 culture of rootage bottom of bottle photo in embodiment 1；
Fig. 3 is that Dwarf Stocks For Apple Trees SH6 is taken root seedling photo in embodiment 1；
Fig. 4 be in embodiment 1 Dwarf Stocks For Apple Trees SH6 tissue cultures take root seedling hole tray growth photo.
Specific embodiment
Following examples are used to illustrate the present invention, but are not limited to the scope of the present invention.Not
In the case of spirit of the invention and essence, the inventive method, step or condition are made
Modification is replaced, and belongs to the scope of the present invention.
If not specializing, technological means used is ripe for those skilled in the art in embodiment
The conventional meanses known.
Embodiment 1
The method of the present invention is tested in In Shunyi District of Beijing Yang Zhen apple tissue culture experiments room,
Dwarf Stocks For Apple Trees aseptic strain is set up, squamous subculture and culture of rootage is carried out, plantlet in vitro Jing of taking root
Field-transplanting has been carried out after domestication, it is specific as follows：
1) foundation of Dwarf Stocks For Apple Trees SH6 aseptic strains
Select the individual plant nutrition branch of Dwarf Stocks For Apple Trees SH6, stem of the clip with terminal bud or lateral bud
Section, explant sterilization is rinsed 30 minutes with running water, and 70% 10～60s of alcohol disinfecting is aseptic
Water is rinsed 2～4 times, and 0.1% mercuric chloride is sterilized 3～10 minutes, aseptic water washing 2～4 times.
Initial culture is carried out, condition of culture is：Temperature 20-28 DEG C, humidity 55-75%, light
According to 14～18 hours/day, intensity of illumination 1800-2200Lx.
Screening polyvinylpyrrolidoneconcentration concentration (PVP-40) concentration, in MS+6-BA
Arrange on the culture medium of 0.4mg/L+IBA 0.3mg/L+ sucrose 25g/L+ agarose 5g/L
The concentration of PVP-40 be 0mg/L, 200mg/L, 400mg/L, 600mg/L, 800mg/L,
1000mg/L, as a result as shown in table 1：
Inhibitory action of the table 1PVP-40 concentration to melting brown rate
| PVP-40 concentration (mg/L) | Pollution rate (%) | Melting brown rate (%) | Survival rate (%) |
| 0 | 32.65±1.36 | 91.03±1.94 | 26.87±1.56 |
| 200 | 35.32±2.65 | 36.37±2.84 | 33.18±2.41 |
| 400 | 34.41±2.07 | 21.68±1.83 | 41.36±2.04 |
| 600 | 36.85±1.67 | 41.83±2.37 | 30.25±1.98 |
| 800 | 37.16±1.83 | 63.85±3.04 | 29.41±2.31 |
| 1000 | 33.82±2.17 | 89.73±2.57 | 27.21±2.09 |
Can see that melting brown rate can be obtained well in 200～400mg/L of PVP-40 by table 1
Suppress, especially during PVP-40 400mg/L, melting brown rate is only (21.68 ± 1.83) %
Finally according to the selection result, the culture medium that Initial culture is adopted is for MS+6-BA
0.1～0.8mg/L+IBA, 0.1～0.6mg/L+PVP-40,200～400mg/L+ sucrose
The culture medium of 25～30g/L+, 5～7g/L of agarose is (i.e. with MS solid mediums as basic culture
Base, the concentration containing 6-benzyl aminopurine is that 0.1～0.8mg/L, the concentration of indolebutyric acid are
0.1～0.6mg/L, polyvinylpyrrolidoneconcentration concentration are that 200～400mg/L, the concentration of sucrose are
25～30g/L, the culture medium that the concentration of agarose is 5～7g/L)
2) the numerous squamous subculture of the expansion of Dwarf Stocks For Apple Trees SH6
Carry out squamous subculture, the Initial culture of Dwarf Stocks For Apple Trees SH6 within 30 days after Initial culture
Test material, adopts culture medium for MS+6-BA 0.1～0.8mg/L+IBA, 0.1～0.6mg/L+
The culture medium of sucrose 25～30g/L+, 5～7g/L of agarose carries out squamous subculture (see Fig. 1), training
Foster condition is：Temperature 20-28 DEG C, humidity 55-75%, hour/day of illumination 14～18, illumination
Intensity 1800-2200Lx, carries out fast breeding；Squamous subculture can be carried out after 30～35 days
Culture of rootage；
3) culture of rootage of Dwarf Stocks For Apple Trees SH6
Screening indolebutyric acid (IBA) concentration, adds respectively 0 on 1/2MS solid mediums
The indolebutyric acid of mg/L, 0.1mg/L, 0.2mg/L ... 1.2mg/L, it is to rooting rate
Affect as shown in table 2.
Impact of the table 2IBA concentration to culture of rootage
As a result show, on culture medium add 0.1～1.2mg/L indolebutyric acid compare without
Rooting rate can be improved, rooting rate can reach when especially the concentration of indolebutyric acid is 0.5～1.0mg/L
To more than 55%, rooting rate reaches more than 80% when the concentration of indolebutyric acid is 0.7～0.9mg/L,
Rooting rate is up to more than 85% when the concentration of indolebutyric acid is 0.7～0.8mg/L, works as indolebutyric acid
Concentration be 0.8mg/L when rooting rate highest.
Therefore, the culture of rootage of Dwarf Stocks For Apple Trees SH6 is concretely comprised the following steps apple dwarf
The squamous subculture material cutting of stock SH6 is simple bud, adopts culture medium for 1/2MS+IBA
The culture medium of 0.1～1.2mg/L+ sucrose 15～25g/L+, 5～7g/L of agarose is (i.e. with 1/2MS
Solid medium is minimal medium, and the concentration containing indolebutyric acid is 0.1～1.2mg/L, sugarcane
Sugar concentration be 15～25g/L, the culture medium that the concentration of agarose is 5～7g/L) taken root
Culture is divided into two stages, light culture stage (see Fig. 2, Fig. 3)：6～12 days, temperature
18-26 DEG C, humidity 55-75%, the optical culture stage：6～10 days, temperature 18-26 DEG C, humidity
55-75%, hour/day of illumination 16～20, intensity of illumination 1800-2200Lx.
4) Dwarf Stocks For Apple Trees SH6 is taken root the hardening and field-transplanting of seedling
When the Dwarf Stocks For Apple Trees SH6 plantlet in vitro root length of culture of rootage reaches 2～3 centimetres, move
Bottle hardening is closed to warmhouse booth 3 days, corkage hardening 3 days then takes out and take root in bottle plantlet in vitro,
Culture medium is washed away, root is carried out using 0.2% carbendazim and is disinfected, be transplanted into hole tray (see
Fig. 4), hole tray filling substrate is peat：Perlite：Vermiculite (1：1：1) mixture, treats
Land for growing field crops is transplanted to after root system development is complete.
The above is only the preferred embodiment of the present invention, it is noted that for this technology neck
For the those of ordinary skill in domain, on the premise of without departing from the technology of the present invention principle, can be with
Some improvements and modifications are made, these improvements and modifications also should be regarded as protection scope of the present invention.
Claims (10)
1. a kind of method of quick breeding apple rootstock SH6, it is characterised in that including as follows
Step：
1) just generation and squamous subculture：By the sterilized sterilization of apple rootstock SH6 containing single leaf
Leaf stem section successively carries out Initial culture and expands numerous subculture training with Initial culture base and subculture medium
Support；
2) culture of rootage：Cut step 1) culture after the stem section containing simple bud, be inoculated into life
Culture of rootage is carried out on root culture medium；
3) hardening and field-transplanting：When the plantlet in vitro root length of culture of rootage reaches 2～3 centimetres,
Carry out hardening and field-transplanting；
The Initial culture base is with MS solid mediums as minimal medium, containing 6- benzyl ammonia
It is 0.1～0.6mg/L, poly- that the concentration of base purine is 0.1～0.8mg/L, the concentration of indolebutyric acid
Vinylpyrrolidone concentration be 200～400mg/L, sucrose concentration be 25～30g/L, agarose
Concentration be 5～7g/L culture medium；
The subculture medium is with MS solid mediums as minimal medium, containing 6- benzyl ammonia
The concentration of base purine be 0.1～0.8mg/L, indolebutyric acid concentration be 0.1～0.6mg/L, sugarcane
The concentration of sugar is 25～30g/L, the culture medium that the concentration of agarose is 5～7g/L；
The root media is with 1/2MS solid mediums as minimal medium, containing Yin
The concentration of diindyl butyric acid be 0.1～1.2mg/L, the concentration of sucrose be 15～25g/L, agarose it is dense
Spend the culture medium for 5～7g/L.
2. the method for claim 1, it is characterised in that the Initial culture base,
Polyvinylpyrrolidoneconcentration concentration is 400mg/L.
3. the method for claim 1, it is characterised in that the root media,
The concentration of indolebutyric acid is 0.5～1.0mg/L.
4. method as claimed in claim 3, it is characterised in that the root media,
The concentration of indolebutyric acid is 0.7～0.9mg/L.
5. method as claimed in claim 4, it is characterised in that the root media,
The concentration of indolebutyric acid is 0.8mg/L.
6. the method for claim 1, it is characterised in that step 1) sterilizing
Sterilize, concrete grammar is：Rinsed 30 minutes with water, the alcohol of concentration expressed in percentage by volume 70% disappears
Poison 10～60 seconds, aseptic water washing 2～4 times, the mercuric chloride sterilization 3～10 of mass percentage concentration 0.1%
Minute, aseptic water washing 2～4 times.
7. the method for claim 1, it is characterised in that the culture of rootage, point
For two stages：
The light culture stage：6～12 days, temperature 18-26 DEG C, humidity 55-75%；
The optical culture stage：6～10 days, temperature 18-26 DEG C, humidity 55-75%, illumination 16～20
Hour/day, intensity of illumination 1800-2200Lx.
8. the method for claim 1, it is characterised in that the hardening, is in temperature
Bottle hardening is closed in the booth of room 3 days, corkage hardening 3 days.
9. the method as described in any one of claim 1, it is characterised in that the land for growing field crops is moved
Plant, to wash away the culture medium of plantlet in vitro of taking root, using the carbendazim of mass percentage concentration 0.2%
Root sterilization is carried out, peat is transplanted into：Perlite：Vermiculite volume ratio is 1：1：1 matrix
In, it is transplanted to land for growing field crops after root system development is complete.
10. the method for quick breeding apple rootstock SH6 described in any one of claim 1-9,
Application in apple is bred.
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Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107371880A (en) * | 2017-07-28 | 2017-11-24 | 济南浩隆生物科技有限公司 | A kind of apple rootstock tissue culturing fast seedling-cultivating method |
| CN107439372A (en) * | 2017-07-11 | 2017-12-08 | 北京市农林科学院 | A kind of quick breeding Dwarf Stocks For Apple Trees C.G 935 method |
| CN107494262A (en) * | 2017-08-28 | 2017-12-22 | 山东省果树研究所 | Tissue culture and rapid propagation method and application and culture medium and preparation method, the disinfection treatment method of apple rootstock bar for apple rootstock |
| CN108124773A (en) * | 2018-01-29 | 2018-06-08 | 宝鸡松良农业科技有限公司 | A kind of method that tissue cultures are carried out using apple stem section |
| CN108157182A (en) * | 2018-02-06 | 2018-06-15 | 山东省烟台市农业科学研究院 | A kind of cultural method for reducing apple Explant browning rate |
| CN108496803A (en) * | 2018-06-19 | 2018-09-07 | 山东省果树研究所 | The efficient rapid propagation method of tissue cultures and culture medium for excellent Dwarf Stocks For Apple Trees ' M9T337 ' |
| CN108782245A (en) * | 2018-06-19 | 2018-11-13 | 山东省果树研究所 | Tissue culture and rapid propagation method and culture medium for the anti-continuous cropping dwarfing rootstock G41 of apple |
| CN109892224A (en) * | 2018-12-24 | 2019-06-18 | 江苏东郁植物科技有限公司 | Apple rootstock " Sprout Free " tissue culture of sprout is commercialized mating system |
| CN111903529A (en) * | 2020-09-17 | 2020-11-10 | 昭通市苹果产业发展中心 | Rapid propagation technology for apple tissue culture seedlings |
| CN112772414A (en) * | 2021-01-08 | 2021-05-11 | 山东农业大学 | Method for improving rapid propagation efficiency of apple rootstock tissue culture seedlings |
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Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107439372A (en) * | 2017-07-11 | 2017-12-08 | 北京市农林科学院 | A kind of quick breeding Dwarf Stocks For Apple Trees C.G 935 method |
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| CN108782245A (en) * | 2018-06-19 | 2018-11-13 | 山东省果树研究所 | Tissue culture and rapid propagation method and culture medium for the anti-continuous cropping dwarfing rootstock G41 of apple |
| CN109892224A (en) * | 2018-12-24 | 2019-06-18 | 江苏东郁植物科技有限公司 | Apple rootstock " Sprout Free " tissue culture of sprout is commercialized mating system |
| CN111903529A (en) * | 2020-09-17 | 2020-11-10 | 昭通市苹果产业发展中心 | Rapid propagation technology for apple tissue culture seedlings |
| CN112772414A (en) * | 2021-01-08 | 2021-05-11 | 山东农业大学 | Method for improving rapid propagation efficiency of apple rootstock tissue culture seedlings |
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